[关键词]
[摘要]
目的 合成人参皂苷Rg1的人工免疫原和包被原,为后续制备人参皂苷Rg1的单克隆抗体及建立相应的免疫分析方法奠定基础。方法 采用NaIO4氧化法分别合成人参皂苷Rg1免疫抗原(Rg1-BSA)和包被抗原(Rg1-PLL);紫外光谱和薄层色谱法鉴定人工抗原偶联是否成功;用间接ELISA方法检测免疫小鼠血清中抗体效价,用间接竞争ELISA检测抗体特异性。结果 经紫外光谱和薄层色谱法检测,人参皂苷Rg1与BSA/PLL偶联成功;免疫小鼠后Rg1-BSA可以使小鼠体内产生抗Rg1抗体;利用合成的偶联物成功建立间接ELISA和间接竞争ELISA方法,测得小鼠血清抗体效价在1∶80 000以上,并且所得抗体能特异性结合人参皂苷Rg1。结论 成功合成了人参皂苷Rg1人工免疫原和包被原,可用于下一步的人参皂苷Rg1单克隆抗体制备及建立相应免疫分析方法。
[Key word]
[Abstract]
Objective To synthesize the artificial antigen of ginsenoside Rg1-bovine serum albumin (Rg1-BSA) and the artificial coated antigen of ginsenoside Rg1-polylysine (Rg1-PLL), and to provide the basis for the preparation of monoclonal antibody (MAb) and the establishment of immunoassay method. Methods Rg1-BSA and Rg1-PLL were synthesized by sodium periodate oxidation method. The characterization of the synthesis was examined by UV spectrometry and TLC method. The titer and specificity of the antibody in serum of immunised mice were detected by indirect enzyme-linked immunosorbent assay (I-ELISA) and indirect competitive enzyme-linked immunosorbent assay (I-CELISA), respectively. Results According to the UV and TLC, the Rg1 was successfully conjugated with BSA and PLL. I-ELISA and IC-ELISA methods were developed using Rg1-PLL. The anti-Rg1 antibody obtained from immunized mice could bind to Rg1 specially and the titer was up to 1∶80 000. Conclusion The artificial immunogen Rg1-BSA and coated antigen Rg1-PLL are successfully synthesized, which could be used to prepare the MAb of Rg1 and establish the immunoassay method.
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[基金项目]
国家自然科学基金资助(30973709,81274043);教育部留学回国人员科研启动基金